>

M-MLV Reverse Transcriptase, RNaseH⁺
(MMLV-RT RNase H⁺, 50,000U)
More Information
CAT #
EBT-1028
QTY
50,000 units (200 unit/㎕)
PRICE (W)
240,000
MANUAL
MSDS
이 분류에 등록된 다른 상품이 없습니다. 확대보기
리뷰 위시 0

상품간략정보 및 구매기능

판매가격 판매중지
배송비결제 주문시 결제

상품의 재고가 부족하여 구매할 수 없습니다.

Description

M-MLV (Moloney Murine Leukemia Virus) Reverse Transcriptase is an RNA-dependent DNA polymerase that can be used in first strand cDNA synthesis using total or poly A⁺ RNA as templates. M-MLV Reverse Transcriptase is purified from a recombinant E.coli clone that carrying pol gene of M-MLV and consists of a single subunit with a molecular weight of 71 kDa. The RNase H activity of M-MLV Reverse Transcriptase is weaker than the commonly used Avian Myeloblastosis Virus (AMV) reverse transcriptase.


Storage Buffer
- 200 unit/μl in 20 mM Tris-HCl, pH7.5, 200 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.01% NP-40, and 50% Glycerol.

Unit Definition
- One unit is the amount of enzyme required for incorporation of 1 nmole of dNTP into acid-insoluble material for 10 min at 37℃.

5x Reaction Buffer
- 250 mM Tris-HCl, pH 8.3, 375 mM KCl, 15 mM MgCl₂, 50 mM DTT.

QC tests
- Activity, SDS-PAGE purity, endonuclease activity test, performance tests.

Storage Condition
- Store at -20℃.

Heat Inactivation
- M-MLV reverse transcriptase can be inactivated by incubation at 90°C for 10 min.

First Strand cDNA Synthesis

1. Add 1 ㎕ of 100 pmol/㎕ primer (random hexamer or oligo d(T)15) or 10 pmol/㎕ of gene specific anti-sense primer per microgram of the total RNA sample in a total volume of ≤10 ㎕ in nuclease-free water.
2. Heat the tube to 70°C for 5 min to melt secondary structure within the RNA template.
3. Cool the tube immediately on ice, then spin briefly to collect the solution at the bottom of the tube.
4. Add the followings to the primer/RNA mix in the order shown.
M-MLV 5X Reaction Buffer 4 ㎕
dNTP mix (2.5 mM each) 4 ㎕
M-MLV RT 200 units 1 ㎕
Add nuclease-free DW to final volume of 20 ㎕
5. Mix gently by flicking the tube, and incubate for 60 min at 37°C or 42°C.
6. To stop reaction, incubate for 5 min at 94°C.

Notes
• The cDNA by reverse transcription can be used for subsequent PCR reactions and for the cDNA library constructions.
• If there is concern about possible RNase contamination in the reaction, Recombinant RNase Inhibitor may be added to the reaction to preserve RNA integrity.

PCR
1. Add the following components to the PCR tubes (for 20 ㎕ total reaction).
10x PCR Buffer 2 ㎕
dNTP mix (2.5 mM each) 1.6 ㎕
Primers (10 pmol/㎕) 0.5 ㎕ each
cDNA by RT reaction 0.1-1 ㎕
Taq (5 unit/㎕) 0.2 ㎕
Add nuclease-free DW to final volume of 20 ㎕

Applications

First strand cDNA synthesis from total RNA or polyA⁺ RNA, Primer extension, RT-PCR 

FAQ
  • 등록된 내용이 없습니다.
Related Products

선택하신 상품이 장바구니에 담겼습니다.

계속구매 장바구니 이동

사이트 정보

회사명 : 회사명 / 대표 : 대표자명
주소 : OO도 OO시 OO구 OO동 123-45
사업자 등록번호 : 123-45-67890
전화 : 02-123-4567 팩스 : 02-123-4568
통신판매업신고번호 : 제 OO구 - 123호
개인정보관리책임자 : 정보책임자명

Copyright © 소유하신 도메인. All rights reserved.